CN111575215A - Heat damage salmonella culture medium and preparation method thereof - Google Patents
Heat damage salmonella culture medium and preparation method thereof Download PDFInfo
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- CN111575215A CN111575215A CN202010509769.1A CN202010509769A CN111575215A CN 111575215 A CN111575215 A CN 111575215A CN 202010509769 A CN202010509769 A CN 202010509769A CN 111575215 A CN111575215 A CN 111575215A
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- 241000607142 Salmonella Species 0.000 title claims abstract description 42
- 239000001963 growth medium Substances 0.000 title claims abstract description 29
- 230000006378 damage Effects 0.000 title claims abstract description 11
- 238000002360 preparation method Methods 0.000 title abstract description 6
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 claims abstract description 40
- ODJQKYXPKWQWNK-UHFFFAOYSA-N 3,3'-Thiobispropanoic acid Chemical compound OC(=O)CCSCCC(O)=O ODJQKYXPKWQWNK-UHFFFAOYSA-N 0.000 claims abstract description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 30
- 239000001888 Peptone Substances 0.000 claims abstract description 24
- 108010080698 Peptones Proteins 0.000 claims abstract description 24
- 235000019319 peptone Nutrition 0.000 claims abstract description 24
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 20
- 239000003833 bile salt Substances 0.000 claims abstract description 20
- 229960001506 brilliant green Drugs 0.000 claims abstract description 20
- HXCILVUBKWANLN-UHFFFAOYSA-N brilliant green cation Chemical compound C1=CC(N(CC)CC)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](CC)CC)C=C1 HXCILVUBKWANLN-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000008103 glucose Substances 0.000 claims abstract description 20
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 20
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 20
- 229940054269 sodium pyruvate Drugs 0.000 claims abstract description 20
- 239000012153 distilled water Substances 0.000 claims abstract description 18
- 102000016938 Catalase Human genes 0.000 claims abstract description 16
- 108010053835 Catalase Proteins 0.000 claims abstract description 16
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims abstract description 16
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims abstract description 13
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims abstract description 13
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims abstract description 12
- 229910001629 magnesium chloride Inorganic materials 0.000 claims abstract description 8
- 208000027418 Wounds and injury Diseases 0.000 claims abstract description 7
- 208000014674 injury Diseases 0.000 claims abstract description 7
- 239000002994 raw material Substances 0.000 claims abstract description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 30
- 238000001816 cooling Methods 0.000 claims description 10
- 238000010438 heat treatment Methods 0.000 claims description 9
- 230000001954 sterilising effect Effects 0.000 claims description 9
- 238000004659 sterilization and disinfection Methods 0.000 claims description 9
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 8
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 3
- 235000013305 food Nutrition 0.000 abstract description 9
- 241000894006 Bacteria Species 0.000 description 13
- 238000001514 detection method Methods 0.000 description 8
- 241000283690 Bos taurus Species 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 239000008223 sterile water Substances 0.000 description 5
- 238000005303 weighing Methods 0.000 description 5
- 239000000872 buffer Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 230000000968 intestinal effect Effects 0.000 description 3
- 230000008439 repair process Effects 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 235000014106 fortified food Nutrition 0.000 description 2
- 230000001965 increasing effect Effects 0.000 description 2
- 231100000225 lethality Toxicity 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000002881 Colic Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- 241000588921 Enterobacteriaceae Species 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 206010039438 Salmonella Infections Diseases 0.000 description 1
- 241000607768 Shigella Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 206010039447 salmonellosis Diseases 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000003685 thermal hair damage Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
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Abstract
The invention relates to the field of food safety, in particular to a heat injury salmonella culture medium and a preparation method thereof, wherein the heat injury salmonella culture medium comprises the following raw materials in parts by mass: 3-15 parts of peptone, 2-10 parts of glucose, 5-30 parts of ox bile salt, 1-15 parts of disodium hydrogen phosphate, 0.1-10 parts of monopotassium phosphate, 0.001-1 part of brilliant green, 0.1-10 parts of 3, 3-thiodipropionic acid (TDPA), 0.01-5 parts of magnesium chloride, 0.01-15 parts of sodium pyruvate, 0.001-1 part of Catalase, 1000 parts of distilled water and pH of 7.0-7.4.
Description
Technical Field
The invention relates to the field of food safety, in particular to a heat injury salmonella culture medium and a preparation method thereof.
Background
Salmonella (Salmonella) is a common food-borne pathogenic bacterium belonging to the family Enterobacteriaceae, the gram-negative bacterium. Typical symptoms after salmonella infection include fever, nausea, vomiting, diarrhea, abdominal cramps, etc., which are serious and even life threatening.
In the food production and processing industry, heat sterilization is always an extremely important sterilization enzyme inactivation mode in the food industry in order to enhance the safety and stability of products. Some bacteria may be damaged due to uneven heating during the thermal processing, and if this factor is ignored during the detection, the result may be distorted. And after thermal processing of food contaminated with salmonella, some of the salmonella may be sublethal by suffering varying degrees of thermal damage. The heat damage bacteria can be repaired under proper conditions, and the physiological and biochemical characteristics and pathogenicity of normal undamaged bacteria are restored. The damaged salmonella in food which can be repaired under proper conditions poses great threat to human health, so that the repair of the heat damaged salmonella and the reduction of the missed detection rate of the salmonella are of great significance.
Disclosure of Invention
Based on the problems, the invention aims to provide a liquid enrichment medium capable of rapidly repairing thermally damaged salmonella in enriched food and a preparation method thereof, which can detect thermally damaged salmonella which cannot be detected according to the existing national standard salmonella detection method for GB4789.4-2016 food safety, reduce the omission ratio and promote the subsequent salmonella detection.
The purpose of the invention is realized by the following technical scheme:
the heat injury salmonella culture medium comprises the following raw materials in parts by weight: 3-15 parts of peptone, 2-10 parts of glucose, 5-30 parts of ox bile salt, 1-15 parts of disodium hydrogen phosphate, 0.1-10 parts of monopotassium phosphate, 0.001-1 part of brilliant green, 0.1-10 parts of 3, 3-thiodipropionic acid (TDPA), 0.01-5 parts of magnesium chloride, 0.01-15 parts of sodium pyruvate, 0.001-1 part of Catalase, 1000 parts of distilled water and pH of 7.0-7.4.
Preferably, the heat injury salmonella culture medium comprises the following raw materials in parts by weight: 5-13 parts of peptone, 3-8 parts of glucose, 8-25 parts of ox bile salt, 5-13 parts of disodium hydrogen phosphate, 1-8 parts of monopotassium phosphate, 0.01-0.8 part of brilliant green, 0.5-8 parts of 3, 3-thiodipropionic acid (TDPA), 0.05-4 parts of magnesium chloride, 0.1-13 parts of sodium pyruvate, 0.005-0.8 part of Catalase, 1000 parts of distilled water and pH of 7.0-7.4.
In one embodiment, the heat-damaged salmonella culture medium comprises the following raw materials in parts by mass: 10 parts of peptone, 5 parts of glucose, 20 parts of ox bile salt, 8 parts of dipotassium phosphate, 2 parts of monopotassium phosphate, 0.015 part of brilliant green, 0.3 part of 3, 3-thiodipropionic acid (TDPA), and Mgcl2.6H20.5 part of O, 4.3 parts of sodium pyruvate and 0.01 part of Catalase.
The invention also provides a preparation method of the heat damage salmonella culture medium, which comprises the following steps:
(1) adding peptone, glucose, ox bile salt, disodium hydrogen phosphate, potassium dihydrogen phosphate, brilliant green, 3, 3-thiodipropionic acid (TDPA) and sodium pyruvate into distilled water according to a certain proportion, heating and dissolving;
(2) after cooling to room temperature, firstly using 10mol/L NaOH and then using 1mol/L NaOH to adjust the pH value to 7.0-7.4;
(3) autoclaving at 115 deg.C for 20 min;
(4) dissolving magnesium chloride and Catalase in a small amount of distilled water, filtering for sterilization, adding into a culture medium which is sterilized under high pressure and cooled to room temperature.
Compared with the prior art, the invention has the following advantages:
1. the present invention selects various formulas and contents thereof, finally determines the nutrient components of peptone and glucose, such as carbon source, nitrogen source, vitamins, growth factor, etc. necessary for the growth of bacteria, and combines a certain amount of ox bile salt, brilliant green and can inhibit gram-positive bacteria and partial gram-negative bacteria. Appropriate amounts of disodium hydrogen phosphate and potassium dihydrogen phosphate were used as buffers to maintain the pH of the medium at 7.0-7.4.
2. 3, 3-thiodipropionic acid (TDPA) is used as an antioxidant, which mainly eliminates free radicals generated in the metabolic process and prevents OH-from having toxic action on injured bacteria. The invention discovers that more than 4 parts of sodium pyruvate has obvious effect on repairing the damaged salmonella; the addition of a small amount of magnesium chloride can compensate the loss of metal ions of the salmonella metabolic enzyme system in the sublethal stateIn combination with catalytic amount of Catalase, H generated in the metabolic process of salmonella is further decomposed2O2。
3. According to the invention, the liquid enrichment medium capable of rapidly repairing the heat-injury salmonella in the enriched food is finally obtained by blending the components and the contents of the formula, and the liquid enrichment medium has a good growth effect and high repairing efficiency. The salmonella with the lethality of 99.99 percent after heat treatment is respectively inoculated into nutrient broth, improved EC broth, intestinal bacteria enrichment broth, Shigella enrichment broth, buffered peptone water, BHI broth and the culture medium, and the result shows that the enrichment speed and the repair effect of the culture medium are obviously superior to those of other culture media.
4. Has better selectivity. The optimal culture temperature of the culture medium is 42 +/-1 ℃, and the culture medium can better inhibit gram-positive bacteria, partial gram-negative bacteria and mixed bacteria at the temperature.
5. And the missing detection rate is reduced. Salmonella with the lethality of 99.99 percent after heat treatment is respectively inoculated into buffer peptone water, BHI broth and the culture medium, and the result shows that the culture medium can detect the salmonella according to the GB4789.4-2016 method, but the salmonella can not be detected in the buffer peptone water.
Detailed Description
The present invention will be further described with reference to the following examples for facilitating understanding of those skilled in the art, and the contents of the examples are not intended to limit the present invention.
The starting materials and reagents used in the present invention are commercially available. Example 1g below represents 1 part.
Example one
Weighing 10.0g of peptone, 5.0g of glucose, 20.0g of bovine bile salt, 8.0g of dipotassium hydrogen phosphate, 2.0g of monopotassium phosphate, 0.015g of brilliant green, 4.6g of 3, 3-thiodipropionic acid (TDPA) and Mgcl2.6H2O0.01g, sodium pyruvate 0.02g, Catalase0.01g, distilled water 1000 ml.
Adding peptone, glucose, ox bile salt, disodium hydrogen phosphate, potassium dihydrogen phosphate, brilliant green, 3, 3-thiodipropionic acid (TDPA), and sodium pyruvate into distilled waterHeating to dissolve, cooling to room temperature, adjusting pH to 7.0-7.4 with 10mol/L NaOH and 1mol/L NaOH, autoclaving at 115 deg.C for 20min, storing at room temperature, and dissolving Mgcl in 5ml sterile water before use2.6H2And O and Catalase are added into the sterilized culture medium after filtration and sterilization and are mixed evenly.
Example two
Weighing 10.0g of peptone, 5.0g of glucose, 20.0g of bovine bile salt, 8.0g of dipotassium hydrogen phosphate, 2.0g of monopotassium phosphate, 0.015g of brilliant green, 0.3g of 3, 3-thiodipropionic acid (TDPA) and Mgcl2.6H2O0.5g, sodium pyruvate 0.02g, Catalase0.01g, distilled water 1000 ml.
Adding peptone, glucose, ox bile salt, disodium hydrogen phosphate, potassium dihydrogen phosphate, brilliant green, 3, 3-thiodipropionic acid (TDPA) and sodium pyruvate into distilled water, heating to dissolve, cooling to room temperature, adjusting pH to 7.0-7.4 with 1mol/L NaOH, autoclaving at 115 deg.C for 20min, storing at room temperature, and respectively dissolving Mgcl in 5ml sterile water before use2.6H2And O and Catalase are added into the sterilized culture medium after filtration and sterilization and are mixed evenly.
EXAMPLE III
Weighing 10.0g of peptone, 5.0g of glucose, 20.0g of bovine bile salt, 8.0g of dipotassium hydrogen phosphate, 2.0g of monopotassium phosphate, 0.015g of brilliant green, 0.3g of 3, 3-thiodipropionic acid (TDPA) and Mgcl2.6H2O0.01g, sodium pyruvate 11.2g, Catalase0.01g, distilled water 1000 ml.
Adding peptone, glucose, ox bile salt, disodium hydrogen phosphate, potassium dihydrogen phosphate, brilliant green, 3, 3-thiodipropionic acid (TDPA) and sodium pyruvate into distilled water, heating to dissolve, cooling to room temperature, adjusting pH to 7.0-7.4 with 1mol/L NaOH, autoclaving at 115 deg.C for 20min, storing at room temperature, and respectively dissolving Mgcl in 5ml sterile water before use2.6H2And O and Catalase are added into the sterilized culture medium after filtration and sterilization and are mixed evenly.
Example 4
Weighing 10.0g of peptone, 5.0g of glucose, 20.0g of bovine bile salt, 8.0g of dipotassium hydrogen phosphate, 2.0g of monopotassium phosphate, 0.015g of brilliant green, 0.3g of 3, 3-thiodipropionic acid (TDPA) and Mgcl2.6H2O0.01g, sodium pyruvate 0.02g, Catalase0.153g and distilled water 1000 ml.
Adding peptone, glucose, ox bile salt, disodium hydrogen phosphate, potassium dihydrogen phosphate, brilliant green, 3, 3-thiodipropionic acid (TDPA) and sodium pyruvate into distilled water, heating to dissolve, cooling to room temperature, adjusting pH to 7.0-7.4 with 1mol/L NaOH, autoclaving at 115 deg.C for 20min, storing at room temperature, and respectively dissolving Mgcl in 5ml sterile water before use2.6H2And O and Catalase are added into the sterilized culture medium after filtration and sterilization and are mixed evenly.
EXAMPLE five
Weighing 10.0g of peptone, 5.0g of glucose, 20.0g of bovine bile salt, 8.0g of dipotassium hydrogen phosphate, 2.0g of monopotassium phosphate, 0.015g of brilliant green, 0.3g of 3, 3-thiodipropionic acid (TDPA) and Mgcl2.6H20.5g of O, 4.3g of sodium pyruvate, 0.01g of Catalase and 1000ml of distilled water.
Adding peptone, glucose, ox bile salt, disodium hydrogen phosphate, potassium dihydrogen phosphate, brilliant green, 3, 3-thiodipropionic acid (TDPA) and sodium pyruvate into distilled water, heating to dissolve, cooling to room temperature, adjusting pH to 7.0-7.4 with 1mol/L NaOH, autoclaving at 115 deg.C for 20min, storing at room temperature, and respectively dissolving Mgcl in 5ml sterile water before use2.6H2And O and Catalase are added into the sterilized culture medium after filtration and sterilization and are mixed evenly.
EXAMPLE six
Salmonella with a heat-induced death rate of 99.99 percent is respectively inoculated into buffer peptone water, BHI broth and the culture medium of the invention and detected according to the GB4789.4-2016 food safety national standard salmonella detection method.
Test examples one to five
Preparing salmonella suspension with 1MFc concentration by using normal saline, and cooling the salmonella suspension after water bath for 5min at 60 ℃ for standby application, wherein the thermal death rate is 99.99%. 10ml of heat-treated bacterial suspension with a concentration of 1MFc was added to 90ml of enriched intestinal bacteria broth, BHI broth, nutrient broth, Shiga broth, mEC broth, EC broth, BPW broth and 90ml of the medium of the present invention, and cultured at 42 ℃. + -. 1 ℃ with a concentration of Mycoleptor measured every 2 hours, and the repair of heat-damaged Salmonella was observed and data was recorded as fast as 3.0M concentration.
| 2h | 4h | 6h | 7h | |
| Test example 1 | 0 | 0.16 | 1.86 | 4.2 |
| Test example two | 0 | 0.18 | 3.98 | 8.2 |
| Test example three | 0 | 0.16 | 2.98 | 6.50 |
| Test example four | 0 | 0.17 | 1.90 | 4.50 |
| Test example five | 0 | 0.63 | 6.20 | 10.8 |
| Enterobacter bacterium-enriching broth | 0 | 0.15 | 1.80 | 3.81 |
| BHI broth | 0 | 0.45 | 5.3 | 6.2 |
| Nutrient broth | 0 | 0.51 | 1.99 | 2.65 |
| Shihe broth | 0 | 0.22 | 2.16 | 2.71 |
| mEC broth | 0 | 0.20 | 1.57 | 1.79 |
| EC broth | 0 | 0.16 | 1.11 | 1.13 |
| BPW broth | 0 | 0.25 | 0.43 | 0.60 |
As can be seen from the table, the bacteria increasing effect is the best in the fifth test example, the fifth test example > the second test example > the third test example > the fourth test example > the first test example > the intestinal bacteria increasing broth > the shiga broth > the nutrient broth > mEC broth > EC broth > BPW broth.
Test example six
Taking 0.4ml of fresh salmonella suspension cultured for 18h in BHI broth, adding into a 9ml physiological saline tube, mixing uniformly, diluting by 10 times, cooling after water bath at 60 ℃ for 5min, repeating the water bath at 60 ℃ for 5min, and cooling for later use. The heat-treated bacterial liquid is used as stock solution, and is diluted by 10 times and then is detected according to the national standard salmonella detection method for GB4789.4-2016 food safety.
Results of plate counting of Salmonella on TSA
Detection results of Heat-treated Salmonella
As can be seen from the table, for salmonella with extremely low content and serious damage, BPW can possibly have omission, and the culture medium has better repairing capability, so that the omission rate is reduced.
Claims (4)
1. The heat injury salmonella culture medium is characterized by comprising the following raw materials in parts by weight: 3-15 parts of peptone, 2-10 parts of glucose, 5-30 parts of ox bile salt, 1-15 parts of disodium hydrogen phosphate, 0.1-10 parts of monopotassium phosphate, 0.001-1 part of brilliant green, 0.1-10 parts of 3, 3-thiodipropionic acid, 0.01-5 parts of magnesium chloride, 0.01-15 parts of sodium pyruvate, 0.001-1 part of Catalase, 1000 parts of distilled water and pH of 7.0-7.4.
2. The culture medium for the heat-damaged salmonella as claimed in claim 1, which comprises the following raw materials in parts by weight: 5-13 parts of peptone, 3-8 parts of glucose, 8-25 parts of ox bile salt, 5-13 parts of disodium hydrogen phosphate, 1-8 parts of monopotassium phosphate, 0.01-0.8 part of brilliant green, 0.5-8 parts of 3, 3-thiodipropionic acid, 0.05-4 parts of magnesium chloride, 0.1-13 parts of sodium pyruvate, 0.005-0.8 part of Catalase, 1000 parts of distilled water and pH of 7.0-7.4.
3. The culture medium for the heat-damaged salmonella as claimed in claim 1, wherein the raw material formula comprises the following components in parts by mass: 10 parts of peptone, 5 parts of glucose, 20 parts of bile salt, 8 parts of dipotassium hydrogen phosphate, 2 parts of monopotassium phosphate, 0.015 part of brilliant green, 0.3 part of 3, 3-thiodipropionic acid and Mgcl2.6H20.5 part of O, 4.3 parts of sodium pyruvate and 0.01 part of Catalase.
4. The method for preparing a culture medium for heat-damaged salmonella as claimed in claims 1 to 3, wherein the method comprises the following steps:
(1) adding peptone, glucose, ox bile salt, disodium hydrogen phosphate, potassium dihydrogen phosphate, brilliant green, 3, 3-thiodipropionic acid and sodium pyruvate into distilled water, and heating for dissolving;
(2) after cooling to room temperature, firstly using 10mol/L NaOH and then using 1mol/L NaOH to adjust the pH value to 7.0-7.4;
(3) autoclaving at 115 deg.C for 20 min;
(4) dissolving magnesium chloride and Catalase in a small amount of distilled water, filtering for sterilization, adding into a culture medium which is sterilized under high pressure and cooled to room temperature.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102864204A (en) * | 2012-09-07 | 2013-01-09 | 四川大学 | Enrichment culture solution for detection of Salmonella in poultry eggs |
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